An integrated approach using advanced bioinformatics tools and targeted gene expression analysis was carried out to evaluate the potential use of seven “housekeeping” genes, commonly employed as internal controls in real-time PCR analysis. Adequate testing of reference gene consistency, is always necessary to validate data from any new experimental conditions, since a unique “housekeeping” suitable for every species, organ, developmental stage and treatment does not exist. Thus, the expression stability of glyceraldehyde-3-phosphate dehydrogenase, elongation factor 1_, actin 11, beta-6 tubulin, polyubiquitin 10, 18S rRNA and 5S rRNA genes were evaluated after exposure to low temperatures and in different organs of Beta vulgaris ssp. vulgaris plantlets. The cDNA sequences were derived from GenBank (NCBI) and TIGR-BvGI (Beta Vulgaris Gene Index), a platform providing high-fidelity tentative consensus (TC), obtained by a reliable and stringent ESTs analysis.
Low temperature effect on housekeeping and sucrose synthase genes expression in sugar beet / Pacifico, D; Moliterni, Vmc; Arru, Laura; Mandolino, G.. - STAMPA. - (2007), pp. .-.. (Intervento presentato al convegno LI Convegno Annuale della Società Italiana di Genetica Agraria tenutosi a Riva del Garda nel 23-26 Sept 2007).
Low temperature effect on housekeeping and sucrose synthase genes expression in sugar beet
ARRU, Laura;
2007
Abstract
An integrated approach using advanced bioinformatics tools and targeted gene expression analysis was carried out to evaluate the potential use of seven “housekeeping” genes, commonly employed as internal controls in real-time PCR analysis. Adequate testing of reference gene consistency, is always necessary to validate data from any new experimental conditions, since a unique “housekeeping” suitable for every species, organ, developmental stage and treatment does not exist. Thus, the expression stability of glyceraldehyde-3-phosphate dehydrogenase, elongation factor 1_, actin 11, beta-6 tubulin, polyubiquitin 10, 18S rRNA and 5S rRNA genes were evaluated after exposure to low temperatures and in different organs of Beta vulgaris ssp. vulgaris plantlets. The cDNA sequences were derived from GenBank (NCBI) and TIGR-BvGI (Beta Vulgaris Gene Index), a platform providing high-fidelity tentative consensus (TC), obtained by a reliable and stringent ESTs analysis.
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